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Veterinaria Medika

ISSN 1979-1305

Vol. 7 / No. 2 / Published : 2014-07

TOC : 14, and page :184 - 193

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Original Article :

Cloning gene fragments non-structural 1 (ns1) of dengue virus subtype 1 (denv-1) as a material candidate of vaccine chimera

Author :

  1. Nur Saidah Said*1
  2. Helen Susilowati*2
  3. Deya Karsari*3
  4. Eryk Hendrianto*4
  5. Mufasirin*5
  6. Fedik Abdul Rantam*6
  1. Mahasiswa Fakultas Kedokteran Gigi
  2. Institut Tropical Disease Universitas Airlangga
  3. Institut Tropical Disease Universitas Airlangga
  4. Institut Tropical Disease Universitas Airlangga
  5. Dosen Fakultas Kedokteran Hewan
  6. Dosen Fakultas Kedokteran Hewan

Abstract :

Dengue infection caused by dengue virus (DENV), which includes family Flaviviridae, which has four serotypes of DENV-1, DENV-2, DENV-3 and DENV-4. Dengue virus has a positive sense RNA genome consisting of three genes encoding structural proteins namely protein C (lipids), the E and M/PRM (glycoprotein) and 7 genes encoding non-structural proteins (NS1, NS2a, NS2b, NS3, NS4a, NS4b and NS5). NS1 protein is a highly conserved glycoprotein that plays an important role in viral replication and maturation, NS1 antigen immunogenicity because it has a high potential, capable of inducing antibodies through binding activity of the complement system. This study aimed to clone and analyze protein NS1 gene encoding the genetic stability of non-structural protein 1 (NS1) of dengue virus subtype 1 (DENV-1) after cloning . The results of this study showed fragments of the gene encoding the NS1 protein of dengue virus subtype 1 (DENV-1) after cloning the DNA plasmid vector (pGEM®-T Easy Vector System) and transformed in E. coli JM109 and then performed the verification testing by PCR followed by electrophoresis showed positive results with the invisibility of amplicon fragments NS1 DENV-1 protein coding genes at 435 bp. After sequencing to look at the stability of the gene fragment encoding the DENV-1 NS1 protein after cloning, there are variations in the composition of which is characterized by the presence of nucleotide insertions, deletions, replacements cloned nucleotide composition compared to the original isolate fragments of the gene encoding the NS1 protein of dengue virus serotype 1 at position initial 1-30 and 425 but the arrangement of positions 31-424 and 426-436 nucleotides stabilized

Keyword :

Dengue NS1, Cloning, Genetic stability,


References :

  1. Alcon, S., A. Talarmin, M. Debruyne, A. Falconar, V. Deubel and M. Flamand., (2002). Enzyme-linked immunosorbent assay spesific to dengue virus type-1 nonstructural protein NS1 reveals circulation of the antigen in the blood during the acute phase of disease in patients experiencing primary or secondary infection. . : J. Clin. Mic. 40: 376-381
  2. Brown, T.A, (2006). Gene cloning and DNA analysis : An introduction. 5th ed. . : Blackwell publishing, Oxford. 233
  3. Kumarasamy, V., A.H. Abdul-Wahab, S.K. Chua, Z. Hassan, Y.K. Chem, M. Mohamad and K.B. Chua, (2007). Evaluation of a commercial dengue NS1 antigen-capture ELISA for laboratory diagnosis of acute dengue virus infection. . : Journal of virological methods: 75-79


   


Archive Article

Cover Media Content

Volume : 7 / No. : 2 / Pub. : 2014-07
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  2. Antibacterial activity of the supernatant of soil isolate bacillus subtilis against aeromonas hydrophila and staphylococcus aureus (in vitro)
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  14. Cloning gene fragments non-structural 1 (ns1) of dengue virus subtype 1 (denv-1) as a material candidate of vaccine chimera
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